P-values for group sequential testing

Group sequential testing procedures are widely employed in long-termclinical trials. We discuss several methods for computing theP-value following a group sequential trial based on differentoutcome-space orderings. We use a criterion defined in thispaper to compare these methods and point out interesting differences.     

Distribution of Cd and Zn in annual xylem rings of young spruce trees [Picea abies (L.) Karst.] grown in contaminated soil

Concentrations and total amounts of Cd and Zn in individual annual xylem increments of 5-year-old spruce trees (Picea abies) were investigated after one growing season in contaminated soils. The plants had been potted in soils amended with different concentrations of Cd and Zn and kept in open air conditions. In the outer xylem rings formed during the treatment period lowest concentrations of Cd and Zn were found. In inner rings concentrations of both elements increased and were highest at the pith. Total amounts of both elements showed reversed distribution patterns with highest values in the outermost rings. The observed increase of total dry matter of the rings from pith towards the outer rings was stronger than the parallel increase in total element amounts in the rings. Thus, concentrations dropped in younger rings. The time of the soil contamination event cannot be inferred from element concentrations in annual rings. The results challenge the concept of monitoring historical trends in trace element pollution using analytical data of tree rings of conifers.     

Moisture-driven changes in the sensitivity of the radial growth of Picea crassifolia to temperature,northeastern Tibetan Plateau

Precipitation is one of the most important climate factors controlling tree growth, yet it is not fully understood how changes in precipitation affect the relationship between growth and temperature. On the northeastern edge of the Tibetan Plateau, nine tree-ring chronologies of Picea crassifolia were developed along a precipitation gradient from semi-arid (mean annual precipitation, 255 mm) to semi-humid (710 mm). We analyze the growth-climate relationships along this precipitation gradient and assess whether these associations are regulated by local precipitation. From 1960 to 2014, temperature increased significantly while precipitation remained stable at the nine sampling sites. The radial growth of P. crassifolia decreased at the semi-arid sites but increased at the semi-humid sites. Growth-temperature relationships gradually changed from negative to positive along the precipitation gradient (from dry to wet sites), particularly during summer. The moist P. crassifolia sites are also characterized by positive correlations with the Palmer Drought Severity Index. The temporal growth-temperature relationships varied significantly among the different spruce sites over the last five decades. Although temperature remains the main factor controlling the growth of P. crassifolia, local precipitation variability is becoming increasingly important. Our findings indicate that considering species distribution areas supports the analyses of the impact of climate change on tree growth.     

Population differentiation in climate sensitivity of resin duct formation during growth resumption in Pinus pinaster

To counteract the effects of herbivores and pathogens, conifers have developed a sophisticated resin-based defensive system. Since defences are costly, trees must continuously accommodate defensive investment throughout plastic responses to environmental stimuli. However, the extent of such responses can differ at the intra-specific level (i.e. genetic variation in plasticity). Here we examined whether and to what extent year-to-year climate fluctuations, an important source of environmental heterogeneity during the trees' life, drive plasticity in defensive allocation of a widespread pine species. Specifically, we quantified interannual variation in resin duct production along a 31-year-period in 174 Pinus pinaster trees of nine range-wide populations grown in two common gardens in Central Spain. We aimed to explore (i) patterns of interannual variation (i.e., temporal plasticity) in resin duct production among populations and sites, (ii) whether such patterns are linked to plastic responses to interannual variation in climate conditions (i.e., climatic plasticity), and (iii) whether plastic responses to climate differ among populations (i.e., genetic variation in plasticity) and sites. We found large interannual plasticity in resin duct production (22.8 % of total variance), with temporal patterns differing among sites and populations. Climate conditions during the early growth period significantly affected the annual differentiation of resin ducts. Particularly, April precipitation had a positive overall effect on resin duct production. Inversely, warmer conditions in April had a negative effect but only in certain populations, which demonstrates genetic variation in climate sensitivity of resin duct formation. Despite significant effects of certain climate variables on annual resin duct production, climate only accounted for a small proportion of the total interannual variation (up to 3.8 % of interannual variation explained by climate factors). This suggests that alternative factors such as trade-offs with growth and temporal variation in biotic and non-climatic abiotic conditions likely contribute to explain interannual fluctuations in defensive investment.     

Activation of Six1 target genes is required for sensory placode formation

In vertebrates, cranial placodes form crucial parts of the sensory nervous system in the head. All cranial placodes arise from a common territory, the preplacodal region, and are identified by the expression of Six1/4 and Eya1/2 genes, which control different aspects of sensory development in invertebrates as well as vertebrates. While So and Eya can induce ectopic eyes in Drosophila, the ability of their vertebrate homologues to induce placodes in non-placodal ectoderm has not been explored. Here we show that Six1 and Eya2 are involved in ectodermal patterning and cooperate to induce preplacodal gene expression, while repressing neural plate and neural crest fates. However, they are not sufficient to induce ectopic sensory placodes in future epidermis. Activation of Six1 target genes is required for expression of preplacodal genes, for normal placode morphology and for placode-specific Pax protein expression. These findings suggest that unlike in the fly where the Pax6 homologue Eyeless acts upstream of Six and Eya, the regulatory relationships between these genes are reversed in early vertebrate placode development.     

Cuticle micromorphology of Saxegothaea (Podocarpaceae)

The cuticle micromorphology of the leaves of the monospecific genus Saxegothaea (Podocarpaceae) was studied by scanning electron microscopy. The external and internal features of the adaxial and abaxial surfaces were characterized. The leaves are hypostomatic. The external adaxial cuticle is rugose with irregular ridges and shallow trench‐like structures that do not correspond to any feature of the inner cuticle surface. The external abaxial cuticle has densely crowded stomata arranged in two bands. The stomata are sunken with pronounced, interrupted Florin rings. Stomatal plugs were not observed. Internally, the adaxial epidermal cells are usually rectangular to square; the abaxial epidermal cells are mainly restricted to the midrib and margins and narrowly rectangular; any among the stomata are irregularly shaped. The stomata are nearly all in direct contact. They show unusual features, including an extra pair of cuticular flanges between the guard cell flanges and those of the lateral subsidiary cells, and ‘bridges’ of lateral subsidiary cell tissue extending polewards above the polar extensions to unite with those at their tips. Neither of these features has been reported previously in Podocarpaceae. The results are discussed in the light of recent phylogenetic studies. It is concluded that, despite its unique cuticular features, Saxegothaea should continue to be regarded as a member of Podocarpaceae. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 58–67.     

Rings without a lord? Enigmatic fossils from the lower Palaeozoic of Bohemia and the Carnic Alps

Fossilized ring‐like structures with enigmatic function and taxonomic affiliation were recovered for the first time from the Upper Ordovician of the Carnic Alps and the Silurian of Bohemia. These rings, already mentioned as minor constituents in previous conodont studies (e.g. Webers 1966, p. 1; Bischoff 1973, p. 147), were reported from the Palaeozoic of several regions in Europe and North America. Originally considered as inwardly accreted adhering discs of a benthic hyolithelminth worm with a phosphatic tubular projection, they were later reinterpreted in relation to a putative crinoid epibiont or even as possible scyphozoans. Despite a long debate, neither the function of the enigmatic Palaeozoic rings nor their taxonomic affiliation has been fully clarified. The studied material, extracted by a standard technique in use for conodonts, consists of 235 elements from 16 stratigraphic levels in the Plöcken Formation (Carnic Alps, Cellon Section; Amorphognathus ordovicicus Biozone, Hirnantian, Ordovician) and in the Kopanina Formation (Bohemia, Mu?lovka Quarry; Polygnathoides siluricus Biozone, Ludfordian, Silurian). To explore whether ring size and shape changed over time, we employed a novel combination of geometric morphometric approaches for outlines with no ‘homologous’ landmarks and showed that only size appreciably varied with an increase of ca. 20%. The emerging data from this study are consistent with the interpretation of the rings as an adhering structure of a benthic organism living on a relatively uniform hard substrate.     

Recent decline in precipitation and tree growth in the eastern Mediterranean

We present evidence of a recent drying in the eastern Mediterranean, based on weather and tree‐ring data for Samos, an island of the eastern Aegean Sea. Rainfall declined rapidly after the late 1970s following trends for the entire Mediterranean and was associated with reduced tree‐ring width in Pinus brutia. The most recent decline led to the lowest annual radial stem increment after the last 100 years (as far as records reach). As moisture availability decreased best correlations of tree growth with rainfall were obtained for progressively longer integration periods (1–2 years in moister periods, 5–6 years during the severe dryness of 20th century's last decades), suggesting increasing dependency in deep soil water. Such long‐term integration periods of tree‐growth responses to precipitation have not been reported before. They may reflect a tree‐rooting pattern adapted to cope with even several successive dry years. In late summer 2000, moisture reserves became exhausted, however, and a substantial fraction of low altitude pines died, including some 80‐year‐old trees, which underlines the exceptional extent this trend had reached. Our findings provide empirical support for Intergovernmental Panel on Climate Change projections derived from global circulation models that the Mediterranean, its eastern basin in particular, should become drier as temperature rises, as was the case in the recent past.     

Structural parameters of amylopectin clusters and semi-crystalline growth rings in wheat starches with different amylose content

Small-angle X-ray scattering (SAXS) and scanning electron microscopy (SEM) were used to investigate the internal structure of wheat starch granules with different amylose content. Different approaches were used for treatment (interpretation) of SAXS data to assess the values of structural parameters of amylopectin clusters and the size of crystalline and amorphous lamella in different wheat starches. The average values of the semi-crystalline growth rings thickness in starches have been determined and the relationship between structural characteristics and thermodynamic melting parameters is discussed.     

Snowmelt and early to mid‐growing season water availability augment tree growth during rapid warming in southern Asian boreal forests

Boreal forests are facing profound changes in their growth environment, including warming‐induced water deficits, extended growing seasons, accelerated snowmelt, and permafrost thaw. The influence of warming on trees varies regionally, but in most boreal forests studied to date, tree growth has been found to be negatively affected by increasing temperatures. Here, we used a network of Pinus sylvestris tree‐ring collections spanning a wide climate gradient the southern end of the boreal forest in Asia to assess their response to climate change for the period 1958–2014. Contrary to findings in other boreal regions, we found that previously negative effects of temperature on tree growth turned positive in the northern portion of the study network after the onset of rapid warming. Trees in the drier portion did not show this reversal in their climatic response during the period of rapid warming. Abundant water availability during the growing season, particularly in the early to mid‐growing season (May–July), is key to the reversal of tree sensitivity to climate. Advancement in the onset of growth appears to allow trees to take advantage of snowmelt water, such that tree growth increases with increasing temperatures during the rapidly warming period. The region's monsoonal climate delivers limited precipitation during the early growing season, and thus snowmelt likely covers the water deficit so trees are less stressed from the onset of earlier growth. Our results indicate that the growth response of P. sylvestris to increasing temperatures strongly related to increased early season water availability. Hence, boreal forests with sufficient water available during crucial parts of the growing season might be more able to withstand or even increase growth during periods of rising temperatures. We suspect that other regions of the boreal forest may be affected by similar dynamics.